EA002809B1 - Intermediate compounds for preparing acyclic nucleosides - Google Patents

Abstract

1. The compound of formula wherein R6 and R7 are C1-7alkyl or benzyl or R6 and R7 together are -CH2-CH2- or -CH2CH2CH2-, -CH2CH2CH2CH2- and R9 is hydrogen or alcohol-protecting group. 2. The compound of Claim 1, wherein R9 is hydrogen. 3. The compound of Claim 1, wherein alcohol-protecting group is a benzyl group. 4. The compound of Claim 1 or 2, wherein each R6 and R7 is ethyl.

Description

The present invention relates to new intermediates for the preparation of derivatives of acyclic nucleosides, which can be used as antiviral drugs used in the fight against herpes and retroviral infections.

The practical use of acyclic nucleosides is limited by their relatively moderate pharmacokinetic properties. In general terms, to solve this problem, a number of approaches using prodrugs were used, the purpose of which was to improve the bioavailability of acyclic nucleosides. One such approach is to prepare ester derivatives, especially aliphatic esters, of one or more hydroxy groups of an acyclic side chain.

EP 165 289 describes a promising anti-herpes agent 9 [4-hydroxy- (2-hydroxymethyl) butyl] guanine, known as H2O. European patent EP 186 640 describes 6-deoxy H2O. In European patent EP 343 133 it is indicated that such compounds, especially the B - (-) enantiomer, also have activity against retroviral infections such as Ηΐν. Various derivatives of H2O, such as phosphonates, aliphatic esters (e.g. diacetate and dipropionate), as well as ethers on the hydroxy groups of the acyclic side chain are disclosed in EP 343 133. This patent also describes methods for the preparation of such derivatives, which include condensation of an acyclic side chain chains at the Ν-position of a typical 6-halogenated purine moiety, or, alternatively, in the closure of the imidazole ring of the pyrimidine or furazano [3,4-b] pyrimidine moiety, or in the closure of the pyrimidine o rings of the imidazole moiety, when the acyclic side chain is already present in the preceding pyrimidine or imidazole moieties, respectively. In accordance with the broadest aspects of each of these methods, the acyclic side chain is pre-substituted, however, special examples show the single-stage diacylation of Н2О with acetic or propionic anhydride and DMF.

Nagpbep et al., 1. Meb. Siesh. 32, 1738 (1989) report the study of a number of lower aliphatic esters of an acyclic nucleoside, 9- [4-hydroxy- (3-hydroxymethyl) butyl] guanine, also known as penciclovir, and its 6-deoxy analogue. Famciclovir, a commercial antiviral agent, is a diacetyl derivative of 6-deoxypenciclovir.

Betasht et al., Ryats. Century. 4 No. 2, 120 (1987) describes lower aliphatic esters of 9 - [(1,3-dihydroxy-2-propoxy) methyl] guanine, also known as ganciclovir. It is noted that dipropionate is the preferred ester.

Lake-Wakaat, et al., Reveals in Αηίίш1стЬ. Adeyk SieshoShag. 33 No. 1, 110-112 (1989) diacetate and dipropionate derivatives of H2O, and also monoacetate and diacetate derivatives of 6-deoxy H2O. It is reported that diacetate and dipropionate derivatives of H2O provide only a slight improvement in bioavailability compared to H2O.

International patent application \ ¥ O 94/24134, published October 27, 1994, discloses aliphatic ester prodrugs of the 6-deoxy-Y-7 analog of ganciclovir, including dipivaloyl, divaleroyl, monovaleroyl, monooleoyl and monostearoyl esters.

Both international patent applications XV O 93/07163, published on April 15, 1993, and \ UO 94/22887, published on October 13, 1994, describe mono-ester derivatives of nucleoside analogues - derivatives of monounsaturated C18 or C20 fatty acids. US Pat. No. 5,216,142, dated June 1, 1993, also describes mono-ester derivatives of fatty acids of nucleoside analogues.

A second approach used to prepare acyclic nucleoside prodrugs involves the preparation of amino acid esters of one or more hydroxy groups of an acyclic side chain. The European patent EP 99 493 describes the amino acid esters of acyclovir, and the European patent application EP 308 065, published March 22, 1989, discloses valine and isoleucine esters of acyclovir.

European patent application EP 375 329, published June 27, 1990, discloses the amino acid ester derivatives of ganciclovir, including divaline, diisoleucine, diglycine and dialanine ester derivatives. The international patent application HUO 95/09855, published April 13, 1995, describes ester derivatives of penciclovir, including monovalin and divalin ester derivatives.

DE 19526163, published February 1, 1996, and also US Pat. No. 5,543,414 of August 6, 1996, describe achiral amino acid esters of ganciclovir.

European patent application EP 694 547, published January 31, 1996, discloses the mono-L-valine ester of ganciclovir and its preparation from divivalgancyclovir.

European patent application EP 654,473, published May 24, 1995, describes various bis-amino acid complex 3 ester derivatives of 9- [1 ', 2'-bis-hydroxymethyl) -cyclopropane-1'-yl] methylguanine.

International Patent Application XVO 95/22330, published August 24, 1995, describes aliphatic esters, amino acid esters, and also acetate / valinate esters of an acyclic nucleoside, 9- [3,3-dihydroxymethyl-4-hydroxy-but-1 -il] guanine. In this work, it is indicated that the bio-applicability decreases when one of the valine esters of the trivalin derivative is replaced by an acetate ester group.

It has been established that H2S ester derivatives with a special combination of amino acid ester and fatty acid ester can significantly improve oral bioavailability compared to the related compound (H2C).

The present invention is the creation of new intermediate compounds

in which Kb and K- are lower C _1-7 alkyl or benzyl, or Kb and B ₇ together are —CH ₂ —CH ₂ - or —CH ₂ CH ₂ CH ₂ -, —CH ₂ CH ₂ CH ₂ CH ₂ -, and K, ₉ is hydrogen or an OH protecting group intended to produce new acyclic nucleosides having antiviral properties that have the following formula I

'' s in which

a) B ₁ represents —C (O) CH (CH (CH ₃ ) ₂ ) XII; or —C (O) CH (CH (CH ₃ ) CH ₂ CH ₃ ) X ₁ H ₂ and B ₂ is —C (O) C ₃ -C ₂₁ saturated or monounsaturated, optionally substituted with OH, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkoxy C _1-6 alkyl, C _1-6 alkanoyl, amino, halo, cyano, azido, mercapto or nitroalkyl;

b) B ₁ is —C (O) C ₃ -C ₂₁ saturated or monounsaturated, optionally substituted with OH, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkoxy C _1-6 alkyl, C _1-6 alkanoyl, amino, halo, cyano, azido, nitro merkaptoili alkyl, and B ₂ represents C (O) CH (CH _{(CH3) 2) 2} CN, or -C (O) CH (CH _(CH3) CH2CH3 ) HH2; and

In ₃ represents OH or H;

and their pharmaceutically acceptable salts.

The beneficial effects of oral administration of the mixed esters of fatty acids and amino acids mentioned above are particularly unexpected in comparison with the corresponding use of esters of fatty acids. Based on the results of the analysis of urinary regeneration (Table 1A) or plasma drug analysis (Table 1B) of rats H2C, it can be concluded that neither mono- or dislocated esters of H2C fatty acids provide any improvement in oral bioavailability compared to related compounds H2C. Indeed, the distearate derivative provides a significantly lower bioavailability than the related compound, which may be due to the undesirable effect of the stearate ester on improving the oral bioavailability of H2C. There are reports that the conversion of one or both hydroxyls in some other acyclic nucleoside analogues to the corresponding valine or diviline ester improves bioavailability. The conversion of Н2С to the corresponding mono- or diwil ester derivatives provides a similar improvement in bioavailability compared to the related compound. Given the fact that fatty acid derivatives of H2C have a negative effect on improving bioavailability, it turned out to be unexpected that mixed ester derivatives of H2C based on amino acid / fatty acid provide improved or comparable oral bioavailability in comparison with such a characteristic of a valine dislike ester derivative of H2C, as evidenced by the data urea-regenerative or plasma drug analyzes, respectively.

Table 1A

Group K ₁ Group K ₂ Bioavailability * Hydrogen Hydrogen 8% Hydrogen Stearoyl 12% Stearoyl Stearoyl one% Felled Hydrogen 29% Felled Felled 36% Felled Stearoyl 56%

See detailed biological example 1 below.

Table 1B

Group K ₁ Group K ₂ Bioavailability * Hydrogen Hydrogen 3.8% Hydrogen Stearoyl 1.9% Stearoyl Stearoyl 0% Felled Hydrogen 31.3% Felled Felled 35% Felled Stearoyl 29%

See biological example 2 below for details.

These acyclic nucleosides derived from the intermediates of the present invention are potent antiviral agents, especially with respect to herpes infections caused by the Wapsa11a virus. Herpes simplex virus type 1 and 2, EpCet-Wagg virus. Negrek type 6 (NNU-b) and type 8 (NNU-8). Such compounds are particularly useful against infection with Wapsa11a virus. such as shingles in the elderly, including post herpes neuralgia, or chicken syphilis in young individuals, when the duration and severity of the disease can be reduced in a few days.

Yeries-Wagg viral infections treatable with the compounds of the invention include infectious mononucleosis / glandular fever that has not previously been treated and which may cause learning failure in adolescents for many months.

The compounds of formula I also have activity against certain retroviral infections, especially such as 81U. H1U-1 and H1U-2, as well as against infections for which a transactivating virus is indicated.

Advantageously, the group H ₃ represents a hydroxy group or its tautomeric form = 0 so that the bulk of the compounds of the invention are naturally occurring guanine, for example, when the side chain is split ίη νίνο. On the other hand, H ₃ may be hydrogen, and in this case, as a rule, they deal with a more soluble b-deoxy derivative, which can oxidize ίη νίνο (for example, using xanthine oxidase) to the guanine form.

The compounds of formula I may be present in racemic form, i.e., they are a mixture of 2K and 28 isomers. However, it is preferable that the compounds of formula I contain at least 70%, preferably at least 90% of the K-form, for example, more than 95% of this form. Most preferably, when the compound of formula I is an enantiomerically pure I-form.

Preferably, the amino acid of the group K ₁ / K ₂ is a derivative of the L-amino acid.

Preferably, the amino acid of the group K ₁ / K ₂ contains an even number of carbon atoms, and mainly represents decanoyl (C ₁₀ ), lauryl (C ₁₂ ), myristoyl (C ₁₄ ), palmitoyl (C ₁₆ ), stearoyl (C ₁₈ ) or eicosanil (C ₂₀ ). Other suitable K ₁ / K ₂ groups include butyryl, hexanoyl, octanoyl or behenoyl (C ₂₂ ). Other suitable K ₁ / K ₂ groups include derivatives of myristoleic, myristelaidine, palmitoleic, palmelaidinoic, n-boktadecenoic, oleic, elaidic, gandoic, erucic or brassidic acids. Monounsaturated fatty acid esters typically have a double bond in the trans configuration, preferably at the ω-6, ω9 or ω-11 position, depending on their length. Preferably, the K ₁ / K ₂ group is a fatty acid derivative containing C9C ₁₇ saturated or p: 9 monounsaturated alkyl.

Saturated or unsaturated fatty acid, or K ₁ / K ₂ may optionally be substituted with the same or different substituents, up to five, selected from the group consisting of substituents such as hydroxy, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ alkoxy C ₁ -C ₆ alkyl, C ₁ -C ₆ alkanoyl, amino, halo, cyano, azido, oxo, mercapto and nitro, etc.

The most preferred compounds of formula I are those in which K ₁ represents —C (O) CHCH (CH ₃ ) ₂ PN ₂ or C (O) CH (CH (CH ₃ ) CH ₂ CH ₃ ) OH ₂ and K ₂ represents a-C (0) C ₉ -C ₁₇ saturated alkyl.

Used in the text, the term lower alkyl refers to alkyl radicals of normal or branched structure, containing 1-7 carbon atoms, including, but not limited to, methyl, ethyl, npropyl, isopropyl, n-butyl, isobutyl, sec. Butyl, tert .- butyl, n-pentyl, 1-methylbutyl, 2,2dimethylbutyl, 2-methylpentyl, 2,2-dimethylpropyl, n-hexyl, etc.

As used in the text, the term “protecting group or Ν-protected” refers to those groups which are intended to protect the terminal Ν-amino acid or peptide, or to protect the amino group from undesirable reactions during synthesis operations.

The traditionally used Ν-protecting groups are described in the book Szheepe Prgoesuye Sgoirk ίη Ogdashs 8up1Ne51k (ίοΐιη \ UPeu & 8opk, No. \ ν Wogk, 1981) and are referred to in this description. Ν-protecting groups include acyl groups such as formyl, acetyl, propionyl, pivaloyl, tert.butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, onitrophenoxyacetyl, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4- bromobenzoyl, 4-nitrobenzoyl and the like; sulfonyl groups such as benzenesulfonyl, p-toluenesulfonyl, and the like; carbamate-forming groups such as benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonylcarboxyloxybenzyloxycarbonyloxycarbonyloxycarbonyloxycarbonyloxycarbonyloxycarbonyloxycarbonyloxycarbonyloxycarbonyloxycarbonyloxycarbonyloxycarbonyloxycarbonyloxycarbonyloxycarbonyloxycarbonyloxycarbonyl 5trimethoxybenzyl-hydroxycarbonyl, 1- (p-biphenylyl) -1-methylethoxycarbonyl, α, α-dimethyl-3,5 dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl, tert-butoxycarbonyl, diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl onyl, allyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, tsiklopentiloksikar7 Bonilla, adamantyloxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl, and the like .; alkyl groups such as benzyl, triphenylmethyl, benzyloxymethyl and the like; silyl groups such as trimethylsilyl and the like. Preferred β-protecting groups are formyl, acetyl, benzoyl, pivaloyl, tert.butylacetyl, phenylsulfonyl, benzyl, tert.butoxycarbonyl (BOC) and benzyloxycarbonyl (Cbx).

As used herein, the term activated ester derivative refers to halides such as acid chlorides, and activated esters include, but are not limited to, anhydrides derived from formic and acetic acids, anhydrides derived from such alkoxycarbonyl halides such as isobutyloxycarbonyl hydroxysuccinimide derivative esters, идhydroxyphthalimide derivative esters, Ν-hydroxybenzotriazole derivative esters, No. hydroxy-5-norbornene-2,3dicarboxamide esters derivatives, 2,4,5-trihlorfenilproizvodnye esters and m. p.

Preferred compounds of formula I include (B) -9- [2- (butyryloxymethyl) -4- (b-isoleucyloxy) butyl] guanine, (B) -9- [2- (4-acetylbutyryloxymethyl) -4- (bisolecyloxy) butyl ] -guanine, (B) -9- [2- (hexanoyloxymethyl) -4- (b-isoleucyloxy) butyl] guanine, (B) -9- [4- (b-isoleucyloxy) -2- (octanoyloxymethyl) butyl] guanine, (B) -9- [4- (b-isoleucyloxy) -2- (decanoyloxymethyl) butyl] guanine, (B) -9- [4- (b-isoleucyloxy) -2- (dodecanoyloxymethyl) butyl] guanine, (B) -9- [4- (L-isoleucyloxy) -2- (tetradecanoyloxymethyl) butyl] guanine, (B) -9- [4- (L-isoleucyloxy) -2- (hexadecanoyloxymethyl) butyl] guanine, (B) -9- [4- (b-isolates cyloxy) -2- (octadecanoyloxymethyl) butyl] guanine, (B) -9- [2- (eicosanoyloxymethyl) -4- (b-isoleucyloxy) butyl] guanine, (B) -9- [2- (docosanoyloxymethyl) - 4- (b-isoleucyloxy) butyl] guanine, (B) -9- [4- (b-isoleucyloxy) -2 - ((9-tetradecenoyl) oxymethyl) butyl] guanine, (B) -9- [2- ((9-hexadecenoyl) oxymethyl) -4 (b-isoleucyloxy) butyl] guanine, (B) -9- [4- (b-isoleucyloxy) -2 - ((6-octadecenoyl) oxymethyl) butyl] guanine, (B ) -9- [4- (L-isoleucyloxy) -2 - ((9 octadecenoyl) oxymethyl) butyl] guanine, (B) -9- [2 - ((11-eicosanoyl) oxymethyl) -4- (L isoleucyloxy ) butyl] -guanine, (B) -9- [2 - ((13-docosenoyl) -oxymethyl) -4- (L isoleucyloxy) butyl] - guanine, (B) -2-amino -9- [2- (butyryloxymethyl) -4- (L isoleucyloxy) butyl] -purine, (B) -2-amino -9- [2- (4-acetyl butyryloxymethyl) -4 - (b-isoleucyloxy) -butyl] purine (B) -2-amino -9- [2- (hexanoyloxymethyl) -4 (b-isoleucyloxy) butyl] purine, (B) -2-amino-9- [4- (B-isoleucyloxy) -2 (octanoyloxymethyl) butyl] purine, (B) -2-amino-9- [4- (b-isoleucyloxy) -2 (decanoyloxymethyl) butyl] -purine, (B) -2-amino -9- [4- (b-isoleucyloxy) -2 (dodecanoyloxymethyl) butyl] purine, (B) -2-amino-9- [4- (b-isoleucyloxy) -2 (tetradecanoyloxymethyl) butyl] purine, ( B) -2-amino-9- [4- (b-isoleucyloxy) -2 (hexadecanoyloxymethyl) butyl] purine, (B) -2 -amino-9- [4- (b-isoleucyloxy) -2 (octadecanoyloxymethyl) -butyl] purine, (B) -2-amino-9- [4- (b-isoleucyloxy) -2 (eicosanoyloxymethyl) -butyl] purine , (B) -2-amino-9- [2- (eicosanoyloxymethyl) 4- (b-isoleucyloxy) butyl] purine, (B) -2-amino-9- [2- (docosanoyloxymethyl) 4- (b- isoleucyloxy) butyl] purine, (B) -2-amino-9- [4- (b-isoleucyloxy) -2 - ((9tetradecenoyl) oxymethyl) butyl] purine, (B) -2-amino-9- [ 2 - ((9-hexadecenoyl) oxymethyl) -4- (b-isoleucyloxy) butyl] purine, (B) -2-amino-9- [4- (b-isoleucyloxy) -2 - ((6-octadecenoyl) hydroxymethyl) butyl] purine, (B) -2-amino-9- [4- (b-isoleucyloxy) -2 - ((9 octadecenoyl) hydroxymethyl) butyl] purine, (B) -2 -amino-9- [2 - ((11-eicosanoyl) oxymethyl) -4- (b-isoleucyloxy) butyl] purine, or (B) -2-amino-9- [2 - ((13-docosenoyl) oxymethyl) -4- (b-isoleucyloxy) butyl] purine, as well as their pharmaceutically acceptable salts.

Other preferred compounds include (B) -9- [2- (butyryloxymethyl) -4- (b-valyloxy) butyl] guanine, (B) -9- [2- (4-acetylbutyryloxymethyl) -4- (bivalyloxy) butyl] guanine, (B) -9- [2- (hexanoyloxymethyl) -4- (b-valyloxy) butyl] guanine, (B) -9- [2- (octanoyloxymethyl) -4- (b-valyloxy) butyl] guanine, (B) -9- [2- (decanoyloxymethyl) -4- (b-valyloxy) butyl] guanine, (B) -9- [2- (dodecanoyloxymethyl) -4- (b-valyloxy) butyl] guanine, (B ) -9- [2- (tetradecanoyloxymethyl) -4- (bivalyloxy) butyl] guanine, (B) -9- [2- (hexadecanoyloxymethyl) -4- (bivalyloxy) butyl] guanine, (B) -9- [2 - (octadecanoyloxymethyl) -4- (bivalyloxy) butyl] gua in, (B) -9- [2- (eicosanoyloxymethyl) -4- (b-valyloxy) butyl] guanine, (B) -9- [2- (eicosanoyloxymethyl) -4- (b-valyloxy) butyl] guanine, (K) -9- [2- (docosanoyloxymethyl) -4- (b-valyloxy) butyl] guanine, (K) -9- [2 - ((9-tetradecenoyl) oxymethyl) -4 (b-valyloxy) butyl] -guanine, (K) -9- [2 - ((9-hexadecenoyl) oxymethyl) -4 (b-valyloxy) butyl] -guanine, (K) -9- [2 - ((6-octadecenoyl) oxymethyl) - 4 (b-valyloxy) butyl] guanine, (K) -9- [2 - ((9-octadecenoyl) oxymethyl) -4 (b-valyloxy) butyl] guanine, (K) -9- [2- ( (11-eicosanoyl) oxymethyl) -4- (bivalyloxy) butyl] guanine, (K) -9- [2 - ((13-docosenoyl) oxymethyl) -4- (bivalyloxy) butyl] guanine, (K) -2- amino 9- [2- (buty riloxymethyl-4- (bivalyloxy) butyl] purine, (K) -2-amino -9- [2- (4-acetylbutyryloxymethyl) -4- (b-valyloxy) butyl] purine, (K) -2-amino -9- [2- (hexanoyloxymethyl) -4 (b-valyloxy) butyl] purine, (K) -2-amino-9- [2- (octanoyloxymethyl) -4 (b-valyloxy) butyl] purine, (K ) -2-amino-9- [2- (decanoyloxymethyl) -4 (b-valyloxy) butyl] purine, (K) -2-amino -9- [2- (dodecanoyloxymethyl) 4- (b-valyloxy) butyl ] -purine, (K) -2-amino-9- [2- (tetradecanoyloxymethyl) -4- (b-valyloxy) butyl] -purine, (K) -2-amino-9- [2- (hexadecanoyloxymethyl) - 4- (b-valyloxy) butyl] purine, (K) -2-amino-9- [2- (octadecanoyloxymethyl) -4- (b-valyloxy) butyl] purine, ( K) -2-amino-9- [2- (eicosanoyloxymethyl) 4- (b-valyloxy) butyl] purine, (K) -2-amino-9- [2- (docosanoyloxymethyl) 4- (b-valyloxy) butyl] purine, (K) -2-amino-9- [2 - ((9-tetradecenoyl) oxymethyl) -4- (b-valyloxy) butyl] purine, (K) -2-amino-9- [ 2 - ((9-hexadecenoyl) oxymethyl) -4- (b-valyloxy) butyl] purine, (K) -2-amino-9- [2 - ((6-octadecenoyl) oxymethyl) -4- (b- valyloxy) butyl] purine, (K) -2-amino-9- [2 - ((9-octadecenoyl) oxymethyl) -4- (b-valyloxy) butyl] purine, (K) -2-amino-9 - [2 - ((11-eicoseonyl) -oxymethyl) -4- (b-valyloxy) -butyl] purine, or (K) -2-amino-9- [2 - ((13-docosenoyl) -oxymethyl) - 4- (b-valyloxy) -butyl] purine;

and their pharmaceutically acceptable salts.

Other preferred compounds of formula I include (K) -9- [4- (butyryloxy) -2- (b-valyloxymethyl) butyl] guanine, (K) -9- [4- (4-acetylbutyryloxy) -2- (b- valyloxymethyl) butyl] guanine, (K) -9- [4- (hexanoyloxy) -2- (b-valyloxymethyl) butyl] guanine, (K) -9- [4- (octanoyloxy) -2- (b-valyloxymethyl) butyl] guanine, (K) -9- [4- (decanoyloxy) -2- (b-valyloxymethyl) butyl] guanine, (K) -9- [4- (dodecanoyloxy) -2- (b-valyloxymethyl) butyl] guanine, (K) -9- [4- (tetradecanoyloxy) -2- (b-valyloxymethyl) butyl] guanine, (K) -9- [4- (hexadecanoyloxy) -2- [b-valyloxymethyl) butyl] guanine, (K) -9- [4- (octadecanoyloxy) -2- (b-valiloksimeti k) butyl] guanine, (K) -9- [4- (eicosanoyloxy) -2- (b-valyloxymethyl) butyl] guanine, (K) -9- [4- (docosanoyloxy) -2- (b-valyloxymethyl) butyl] guanine, (K) -9- [4 - ((9-tetradecenoyl) oxy) -2- (L-valyloxymethyl) butyl] guanine, (K) -9- [4 - ((9-hexadecenoyl) oxy ) -2- (b-valyloxymethyl) butyl] guanine, (K) -9- [4 - ((6-octadecenoyl) oxy) -2- (b-valyloxymethyl) butyl] guanine, (K) -9- [4 - ((9-octadecenoyl) oxy) -2- (b-valyloxymethyl) butyl] guanine, (K) -9- [4 - ((11-eicosenoyl) oxy) -2- (b-valyloxymethyl) butyl ] guanine, (K) -9- [4 - ((13-docosenoyl) -oxy) -2- (b-valyloxymethyl) butyl] guanine, (K) -2-amino -9- [4- (butyryloxy) -2 - (b-valiloximethyl) butyl] p rin, (K) -2-amino-9- [4- (4-acetylbutyryloxy) -2 (L-valyloxymethyl) -butyl] purine, (K) -2-amino-9- [4- (hexanoyloxy) -2 - (b-valyloxy) butyl] purine, (K) -2-amino-9- [4- (octanoyloxy) -2- (b-valyloxymethyl) butyl] purine, (K) -2-amino-9- [4 - (decanoyloxy) -2- (b-valyloxymethyl) butyl] purine, (K) -2-amino-9- [4- (dodecanoyloxy) -2- (bvalyloxymethyl) butyl] purine, (K) -2 -amino -9- [4- (tetradecanoyloxy) -2 (b-valyloxymethyl) butyl] -purine, (K) -2-amino-9- [4- (hexadecanoyloxy) -2 (b-valyloxymethyl) butyl] -purine, ( K) -2-amino-9- [4- (octadecanoyloxy) -2- (bivaloxylmethyl) butyl] purine, (K) -2-amino -9- [4- (eicosanoyloxy) -2- (bivaloc imethyl] butyl purine, (K) -2-amino -9- [4- (up to kosanilo xi) -2- (bivalixymethyl) butyl] purine, (K) -2-amino-9- [4 - (( 9-tetradecenoyl) oxy) 2- (b-valyloxymethyl) butyl] purine, (K) -2-amino-9- [4 - ((9-hexadecenoyl) oxy) 2- (b-valyloxymethyl) butyl] purine , (K) -2-amino-9- [4 - ((6-octadecenoyl) oxy) 2- (b-valyloxymethyl) butyl] purine, (K) -2-amino-9- [4 - ((9 -octadecenoyl) oxy) 2- (b-valyloxymethyl) -butyl] purine, (K) -2-amino-9- [4 - ((11-eicosenoyl) oxy) -2 (b-valyloxy) butyl] -purine, (K) -2-amino-9- [4 - ((13-docoseonyl) oxymethyl) -2 - (b-valyloxy) butyl] purine, or pharmaceutically acceptable salts of these compounds.

The compounds of formula I can form salts. Suitable pharmaceutically acceptable salts of the compounds of formula I include salts of organic acids, mainly carboxylic acids, including, but not limited to, acetate, trifluoroacetate, lactate, gluconate, citrate, tartrate, maleate, malate, pantothenate, isoethionate, adipate, alginate, aspartate , benzoate, butyrate, digluconate, cyclo pentanate, glucoheptane, glycerophosphate, oxalate, heptanoate, hexanoate, fumarate, nicotinate, palmoate, pectinate, 3-phenylpropionate, picrate, pivalate, propionate, tartrate, lactobate, lactobate, lactobate, camphor tsinat such salts of organic sulfonic acids such as methanesulfonate, ethanesulfonate, 2-hydroxyethanesulfonate, camphorsulfonate, 2-naphthalenesulfonate, benzenesulfonate, p-chlorobenzenesulfonate and p-toluenesulfonate; as well as salts of inorganic acids, such as hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, hemi sulfate, thiocyanate, persulfate, as well as salts of phosphoric and sulfonic acids. It is most convenient to use hydrochloric acid salts.

The compounds of formula I can be isolated as hydrates, in crystalline form, preferably in the form of homogeneous crystals, in which case an additional aspect of the present invention provides compounds of formula I in a substantially pure crystalline form containing> 70%, preferably> 90% of a homogeneous crystalline material for example> 95% homogeneous crystalline material.

The compounds of formula I are particularly suitable for oral administration, however, they can be administered rectally, vaginally, nasally, topically, transdermally or parenterally, for example intramuscularly, intravenously or epidurally. The compounds of the invention can be used as such, for example, in the form of capsules, or, as a rule, together with a pharmaceutically acceptable carrier or diluent.

Oral formulations are most conveniently formulated in unit dosage forms such as capsules or tablets using conventional carriers or binders such as magnesium stearate, chalk, starch, lactose, wax, gum or gelatin. Liposomes, or synthetic or natural polymers such as HPMC or RUR, can be used to create sustained release formulations. On the other hand, such formulations can be made in the form of nasal or eye drops, syrup, gel or cream containing a solution, suspension, emulsion, oil-in-water or water-in-oil preparation, in such conventional excipients as water, saline, ethanol, vegetable oil or glycerin, optionally in the presence of a fragrance and / or preservative and / or emulsifier.

The compounds of formula I can be used with a daily dosage in the range of 0.1-200 mg / kg / day, preferably 0.5-100 mg / kg / day, more preferably 10-50 mg / kg / day, for example 10-25 mg / kg / day. A typical dosage for a normal adult can be 50-500 mg, for example, 300 mg, which is used once or twice a day in the case of herpes and 2-10 times in the case of ShU infections.

In accordance with the usual rules of antiviral therapy, the compounds can be used in combination with other antiviral agents such as acyclovir, valciclovir, penciclovir, famciclovir, ganciclovir and their prodrugs, cidofovir, foscarnet and similar drugs prescribed for the treatment of herpes, as well as ΑΖΤ, bq bbs, 64T, 3TS, foscarnet, ritonavir, indinavir, saquinavir, delaviridine. Uyeh UH 478, Adoigoy ΑΟ1343 and other drugs prescribed for the treatment of retroviral infections.

Scheme C illustrates the preparation of an intermediate compound according to the invention (compound 11 in the scheme) and its subsequent conversion into a biologically active compound of formula I.

Scheme C

TO,

Formula I

According to Scheme C, malonate 1 (K ₄ and K ₅ are lower alkyl, benzyl or the like) are alkylated by reaction with about 0.5 to 2.0 molar equivalents of acetal 2 (Kb and K ₇ are lower alkyl, benzyl and the like, or Kb and K ₇ together represent —CH ₂ CH ₂ - or —CH ₂ CH ₂ Sn ₂ -, or

CH2CH2CH2CH2-, and Χι represents a leaving group (e.g. C1, Br or I, or a sulfonate such as methanesulfonate, triflate, ptoluenesulfonate, benzenesulfonate, etc.) in the presence of 0.5-2.0 molar equivalents of base (e.g. , potassium tert.-butylate, or sodium ethylate, or ΝαΗ or KH, etc.) in an inert solvent (for example, such as DMF, THF, dioxane, dioxolane, Ν-methylpyrrolidone, etc.) at a temperature of the range of -40-190 ° C. to obtain alkyl malonate 3.

As a result of reduction 3, with a ratio of 0.5-4.0 molar equivalents of ester to alcohol reducing agent (for example, such as NIV ₄ , Ca (VN ₄ ) ₂ or N ₄ , N ₁ H ₄ , etc.) in an inert solvent (for example, THF, methyl tert.butyl ether or tert.-ViOH, or the like) at a temperature in the range from -20 to about 10 ° C, diol 4 was obtained. As a result of enzymatic esterification 4 by reaction 1,0-20,0 with molar equivalents of a vinyl ester 5 (K. ₈ represents a C ₃ -C ₂₁ saturated or monounsaturated, optionally substituted al il) in the presence of lipase (e.g., P8-30 lipase, PPb lipase, CCL lipase, etc.) or phospholipase (e.g. phospholipase E, etc.), the desired stereoisomer of ester 6 is obtained. Such a reaction can be carried out in the absence of solvent or in the presence of an inert solvent (for example, methyl tert-butyl ether, toluene or hexane, etc.). The reaction was carried out at a temperature in the range from -20 to about 80 ° C.

The alcohol substituent of compound 6 is converted to a leaving group (for example, such as halogen or sulfonate) by reaction with a halogenating agent (for example, such as No. 8 / P (Pu) ₃ or Νί'8 / Ρ (Ρ1ι) _3. Either POC1 ₃ or ΝίΈ / Ρ (Ρ1ι) ₃ / ΝαΙ in an acetone or similar solvent) in an inert solvent (such as methylene chloride, toluene, ethyl acetate, etc.) or by reaction with 0.82.0 molar equivalents of sulfonyl halide (for example, such as benzenesulfonyl chloride, toluenesulfonyl chloride or methane sulfonyl chloride, etc.) in the presence of 1.0-4.0 molar equivalents of a base (for example, such as triethylamine, potassium carbonate, pyridine, dimethylaminopyridine or ethyldiisopropylamine, etc.) in an inert solvent (for example, such as methylene chloride, toluene, ethyl acetate, pyridine, methyl tert .-butyl ether and the like) at a temperature in the range of 25 ° C. to about 100 ° C. to give the ester 7 (X ₂ represents a halogen or sulfonate leaving group).

According to the reaction of compound 7 with 0.9-2.0 molar equivalents of 2-amino-4-chloropurine 8 in the presence of 1.0-6.0 molar equivalents of a base (for example, such as potassium carbonate, ΝαΗ, KN, NaOH, KOH lithium diisopropylamide, etc.) in an inert solvent (for example, such as DMF, THF, acetonitrile, Ν-methylpyrrolidone ethanol, etc.) at a temperature in the range from -25 ° C to about 140 ° C received substituted purine 9.

Alternatively, by combining (for example, using the P (Pb) ₃ / diethyl azidocarboxylate system) alcohol 6 with 2-amino-4-chloropurine 8, compound 9 is obtained.

As a result of reaction of 9 with 2.0-20 molar equivalents of alcohol K. ₉ OH (K. ₉ is an alcohol protecting group such as benzyl, etc.) in the presence of 1.0-6.0 molar equivalents of a base (for example, such as potassium tert.-butylate, potassium carbonate, NN KN, lithium diisopropylamide, etc.) in an inert solvent (for example, such as THF, DMF, etc.), at a temperature of -25 ° С to about 150 ° C. alcohol 10 was obtained.

By removing the alcohol protecting group P _{9 of} compound 10 (for example, by catalytic hydrogenation in an inert solvent such as ethanol, benzyl alcohol, methanol, THF and the like in the presence of a hydrogenation catalyst such as Pb / C or Pb (OH ) ₂ , etc.), substituted guanine 11 was obtained.

As a result of esterification of compound 11 by reaction with a) 0.8-2.0 molar equivalents of K. ₁₀ COOH and a crosslinking agent (for example, DSS / DMAR), etc. in an inert solvent (e.g. THF, DMF, etc.) or b) with 0.82.0 molar equivalents of the activated derivative P | ₀ COOH (for example, such as acid chloride, or Ν-hydroxysuccinimide ester, or K.uC (O) OC (O) K.1 ₀ , etc.) in the presence of 0-3.0 molar equivalents of base (for example such as pyridine, triethylamine, ethyldiisopropylamine, TLD, potassium carbonate, etc.) in an inert solvent (for example, such as methylene chloride, THF, pyridine, acetonitrile, DMF, etc.) at a temperature of -25 ° С to about 100 ° C., an ester of 1 2 was obtained.

The acetal substituent of compound 12 was removed and the resulting aldehyde was reduced by reacting compound 12 with 0.1-10.0 molar equivalents of an acid (such as triflic acid, HC1, acetic acid, sulfuric acid, etc.) in an inert solvent ( for example, such as THF / H ₂ O, methylene chloride / H ₂ O, ethyl acetate / H ₂ O, ethanol / H ₂ O, methanol / H ₂ O, etc.) at a temperature in the range from -25 ° C to, approximately 100 ° C. 0.1-10.0 molar equivalents of a base (for example, such as sodium bicarbonate, potassium carbonate, triethylamine, pyridine, KOH, etc.), an additional amount of an inert solvent (for example, such as THF, methylene chloride) were added to the crude mixture. , ethyl acetate, methyl tert.-butyl ether, isopropanol, etc.), as well as from 0.3 to 5.0 molar equivalents of an aldehyde reducing agent (for example, such as sodium borohydride, KaN / H ₂ , etc. .), at a temperature in the range from -25 to 100 ° C, to obtain alcohol 13.

As a result of reaction 13 with 0.8-3.0 molar equivalents of the защищ-protected amino acid P1YCHS (K ₁₁ ) COOH or its activated derivative (P ₁ represents a группу protecting group, and Pc represents isopropyl or isobutyl) in an inert solvent (for example such as THF, dioxane, dioxolane, DMF, methylene chloride, etc.) at a temperature of 25-100 ° C, received alcohol 14.

By removing the сн-protecting group of compound 14, a compound of the present invention of formula I is obtained in which K ₃ is —OH.

Alternatively, compound 13 may be reacted with a symmetric anhydride derived from Ρ | ΝΗί.Ή (Ρ ||) ίΌΟΗ (for example, P ^ CH ^ EOOOO-aOOCH ^ OMNP!) To give compound 1, in which K ₃ represents IT.

Example 1. Obtaining (K) -9- [4-diethoxy-2 (hydroxymethyl) butyl] guanidine he

. her- / oa

a) Obtaining ethyl 4,4-diethoxy-2ethoxycarbonylbutyrate

EY ₂ Cx_, CO ₂ E1 ^ OE (OE1

Potassium tert-butylate (141.8 g, 1.11 eq.) Was dissolved in dry DMF (1 L). Diethyl malonate (266 ml, 1.54 equiv.) Was added over 5 minutes. Bromoacetaldehyde diethyl acetal (172 ml, 1.14 mol) was added over 5 minutes. The mixture was heated to 120 ° C (internal temperature) and stirred at 120 ° C for 5 hours. The resulting mixture was allowed to cool to room temperature, poured into water (5 L) and methyl tert was extracted. -butyl ether (MTBE, 3 x 600 ml). The organic solution was dried over MgSO ₄ , filtered, concentrated, and distilled (0.5 mm, 95–140 ° С) to obtain the desired diester (244 g, 78%) as a colorless oil.

Ή NMR spectrum (СОС1 ₃ ) δ 1.19 (t, 6Н), 1.28 (t, 6Н), 2.22 (dd, 2Н), 3.49 (m, 2Н), 3.51 (t, 1H), 3.65 (m, 2H), 4.20 (quadruplet, 4H), 4.54 (t, 1H).

B) Preparation of 4,4-diethoxy-2- (hydroxymethyl) -butanol

BUT ^ X ^ OH

Ι ^, ΟΕΙ

ΟΕ "

L1BH ₄ (trade solution, 2M in THF, 22.5 ml) and the product from step a) of Example 1 (5 g in 15 ml of THF, 18.1 mmol) were combined with each other, heated to 60 ° C and stirred at 60 ° C for 4 hours. The reaction mixture was allowed to cool to room temperature and the reaction vessel was placed in a cold water bath. Then, triethanolamine (5.97 ml, 1 equiv.) Was added at such a rate that the temperature of the reaction mixture was maintained in the range of 2025 ° C. Brine (17.5 ml) was added at such a rate that gas evolution was controlled and the resulting mixture was stirred for 45 minutes at room temperature. The stratification of the system occurred and the organic layer was washed with brine (2 x 15 ml). The combined brine washes were extracted with MTBE (methyl tert.-butyl ether, 3 x 20 ml). The combined organic extracts were evaporated and the residue was dissolved in MTBE (50 ml) and washed with brine (25 ml). The brine layer was back-extracted with MTBE (3 x 25 ml). The combined organic extracts were dried over # 8O ₊ filtered and concentrated to give the desired diol (3.36 g,

15.5 mmol, 97%) as a colorless oil.

Spectrum ^! H NMR (SPS1 ₃ ) δ 1.22 (t, 6H), 1.73 (dd, 2H), 1.92 (m, 1H), 2.67 (broad s, 2H), 3.52 (m, 2H), 3.69 (m, 2H), 3.72 (m, 4H), 4.62 (t, 1H).

c) Preparation of (2K) -2-acetoxymethyl-4,4-diethoxybutanol

The product from step b) of Example 1 (3.84 g, 20 mmol) was loaded into a 10-ml 1-neck round-bottom flask, then vinyl acetate (2.6 g, 30 mmol) was added, and finally P8 30 lipase (69 mg obtained from Atapo, Lomber, ΙΠίηοίδ). The mixture was stirred at ambient temperature for 16 hours. The progress of the reaction was closely monitored by TLC (2/1 hexane-EJAc; staining with Ce ₂ (8O ₄ ) ₃ and carbonization on a hot plate; Γ.ί. diol 0.1, monoacetate -0.3, bis-acetate -0.75). The reaction mixture was diluted with CH2C12 and filtered through a 5 micron filter. The filter was washed with additional CH ₂ Cl ₂ . Then the filtrate was concentrated in vacuo to obtain the desired product.

b) Preparation of (28) -2-acetoxymethyl-4,4-diethoxybutyltoluene sulfonate

HER— (

ΟΕΙ

In a 100-ml 1-necked round-bottom flask equipped with a magnetic stirrer and a membrane, the crude product from step c) of Example 1 (4.62 g, 19 mmol), dry CH ₂ Cl ₂ (20 ml) and Εΐ _{3 were} charged under nitrogen atmosphere. Ν (5.62 ml, 40 mmol). Tosyl chloride (4.76 g, 25 mmol) was added to the resulting solution.

The resulting mixture was stirred at ambient temperature for 4 hours. H ₂ O (0.27 g, 15 mmol) was charged and the mixture was stirred vigorously for 4 hours. The reaction mixture was diluted with 80 ml of EAAc and 50 ml of H ₂ O and aqueous the layer was separated. To the organic layer was added 75 ml of a 5% aqueous KH2RO4 solution. After mixing and separation of the layers, the aqueous layer was discarded. The organic layer was washed with 50 ml of saturated solution No. HCO ₃ . dried over Na ₂ 8 O ₄ , filtered and concentrated in vacuo to constant weight, resulting in 7.40 g of the desired product.

Spectrum '11 NMR (СОС1 ₃ ) δ 1.17 (t, 6Н), 1.62 (m, 2Н), 1.94 (s, 3Н), 2.19 (m, 1Н), 2.45 (s , 3H), 3.42 (m, 2H), 3.6 (m, 2H), 4.03 (m, 4H), 4.51 (t, 1H), 7.36 (d, 2H), 7 79 (d, 2H).

e) Obtaining α

HER— /

OE1

The product from step b) of Example 1 (3.88 g, 10 mmol), anhydrous DMF (20 ml), 2 amino-4-chloropurine (2.125 g of 12.5 mmol) and K2CO3 (1 ml) were loaded into a 50-ml 1-neck round-bottom flask. 4.83 g). The resulting suspension was stirred at 40 ° C in an atmosphere of Ν ₂ for 20 hours. The mixture was concentrated to remove most of the DMF on a rotary evaporator. The residue was diluted with ELS (50 ml) and H ₂ O (50 ml). The reaction mixture was transferred to a separatory funnel, shaken and the aqueous layer was separated. The aqueous layer was extracted with E1OLs (25 ml). The organic layers were combined and washed with 5% KH2PO4 (75 ml). The organic layer was separated and washed with H2O (75 ml), brine (75 ml), dried over Na ₂ 8O ₄ , filtered and concentrated in vacuo to give 3.95 g of crude product. This crude product was suspended with 40 ml of methyl tert.-butyl ether. The resulting mixture was stirred overnight at 4 ° C and filtered. The filtrate was concentrated to give 3.35 g of the product as an oil (containing 2.6 g of the expected product in accordance with HPLC analysis).

300 MHz '11 NMR (SES1 ₃ ) δ 1.19 (m, 6H), 1.69 (2H), 1.79 (s, 1H), 2.03 (s, 3H), 2.52 (m, 1H), 3.48 (m, 2H), 3.62 (m, 2H), 4.04 (m, 2H), 4.16 (m, 2H), 4.61 (t, 1H), 5, 12 (broad s, 2H); 7.81 (s, 1H).

ί) Receiving

ORP

Ek 'Υ' ⁴ he

EYU / OE1 (Vp = benzyl)

Benzyl alcohol (136 ml) was charged into a 500 mL 1-necked round-bottom flask, the mixture was cooled to 0 ° C, after which KO-tert-Vi (36 g, 321 mmol) was added portionwise. The mixture was allowed to warm to 40 ° C. and the mixture was stirred for 20 minutes. The crude product from step e) of Example 1 (24.7 g, 64.2 mmol) dissolved in 25 ml of anhydrous THF and benzyl alcohol (30 ml) was added to the resulting mixture at 0 ° C. The temperature was allowed to slowly rise to 8 ° C in two hours. The reaction mixture was poured into 500 ml of ice and extracted with 500 ml of MTBE. The organic layer was washed with 250 ml of brine, dried over Na ₂ 8 O ₄ , filtered and concentrated in vacuo to give 193 g of a solution of the target product in benzyl alcohol. In accordance with HPLC analysis, it was found that the solution contained 25.96 g of the target product.

300 MHz '11 NMR (SES1 ₃ ) δ 1.22 (m, 6H), 1.55 (2H), 2.18 (m, 1H), 3.15 (m, 1H), 3.40 (m, 1H); 3.51 (m, 2H), 3.70 (m, 2H), 4.25 (m, 2H), 4.63 (t, 1H), 4.90 (broad s, 2H), 5.25 ( m, 1H), 5.58 (s, 2H), 7.35 (m, 3H), 7.51 (m, 2H), 7.72 (s, 1H).

M8 = (M + H) + = 416 (C1).

d) Obtaining the connection specified in the header.

The crude product from step ί) of example 1 (9.65 g of a solution in benzyl alcohol containing 1.30 g, 3.13 mmol of the product of step ί) of example 14 (dissolved in absolute EH ( 20 ml). To the solution was added 0.45 g of 10% Pb / C suspended in 5 ml of absolute E1OH. The reaction flask was evacuated and H2 was filled three times with hydrogen. The pressure in the reaction flask was H2 at 1 atm and the reaction mixture stirred overnight.The reaction mixture was filtered through a pad of diatomaceous earth I deletion Rb / C. The volatiles were removed in vacuo. The residue was mixed with 25 ml of isopropyl acetate and then concentrated in vacuo. The residue was diluted EYuLs (10 ml) was added the title product seeded, heated to reflux and then was added CH ₃ CN (2 ml) and MTBE (35 ml), then the resulting mixture was stirred for 30 minutes, the precipitate was filtered off and dried to constant weight to obtain 600 mg of the desired product.

300 MHz Ί1 NMR (b ₂ -EM8O) δ 1.16 (m, 6H), 1.45 (m, 1H), 1.61 (m, 1H), 2.16 (m, 1H), 3.45 (m, 2H), 3.40 (m, 1H), 3.62 (m, 2H), 4.02 (m, 2H), 4.53 (t, 1H), 4.85 (t, 1H) 6.55 (broad s, 1H); 7.75 (s, 1H); M8 = (M + H) + = 416 (C1).

In a similar way, starting from the corresponding 4,4-dibenzoxy-2-ethoxycarbonyl butyrate and 4-alkylenedioxy-2-ethoxycarbonyl butyrate, the inventive intermediates are obtained in which He and P- are benzyl or together form —CH2 — CH2—, —CH2CH2CH2— groups , -CH2-CH2-CH2-CH2-.

Example 2. Obtaining (H) -9- [4-hydroxy-2 (stearoyloxymethyl) butyl] guanine.

a) Obtaining OH, η / γΎν 9

Y ^ O ^L (CH ₈ ) ₁₆ CH 3 b! O- /

OE1

(B) -9- [4-diethoxy-2- (hydroxymethyl) butyl] guadine obtained in Example 1 (0.650 g, 2.0 mmol), pyridine (4 ml) were charged into a 1-necked round-bottom flask with a capacity of 25 ml CH ₂ Cl ₂ (2 ml), DMAP (10 mg). The resulting mixture was cooled to −5 ° C. and stearoyl chloride (790 mg, 2.6 mmol) dissolved in CH ₂ Cl ₂ (0.5 ml) was added over 5 minutes. The resulting mixture was stirred for 16 hours at -5 ° C. Absolute ethanol (0.138 g, 3.0 mmol) was added and the mixture was stirred for another 1 hour. Then the reaction mixture was concentrated in vacuo. Toluene (30 ml) was added to the residue, and then the mixture was concentrated in vacuo. Toluene (30 ml) was again added to the obtained residue, and the mixture was concentrated in vacuo. To the resulting residue was added 1% KH ₂ PO ₄ (25 ml) and this mixture was extracted with CH ₂ Cl ₂ (60 ml). The organic layer was separated and dried over Ia ₂ 8o _4, filtered and concentrated in vacuo to a constant weight of 1.65 g crude product was chromatographed on 40 g of ₂ occupies 8, eluting with CH ₂ C1 ₂ - EtOH 95/5, resulting in 367 mg of the expected product.

300 MHz Ή NMR (СОС1 ₃ ) δ 0.89 (t, 3Н), 1.26 (m, 30Н), 1.65 (m, 3Н), 2.32 (m, 1Н), 3.45 (m , 1H), 3.60 (m, 2H), 4.08 (m, 2H), 4.60 (m, 1H), 6.0 (broad s, 2H), 7.53 (s, 1H).

B) Preparation of the compound indicated in the table of contents

N about

I. Ϊ: O (CH2) ₁₆ CH ₃

BUT - /.

The product from step a) of Example 2 (0.234 g, 0.394 mmol) dissolved in THF (1.7 ml) was charged into a 25-mL 1-neck round bottom flask. Triflic acid (0.108 g) in 180 mg H ₂ O was added to this solution. The resulting mixture was stirred overnight at room temperature. To the reaction mixture was added a saturated solution of IAHCO ₃ (10 ml), THF (5 ml), CH ₂ Cl ₂ (2 ml) and IAB ₄ (0.10 g). The mixture was stirred for 30 minutes. A 5% KH2RO4 solution (30 ml) was added to the reaction mixture. The resulting mixture was extracted with 2 x 15 ml of CH ₂ Cl ₂ . The organic layers were combined and dried over IA ₂ 8O ₄ , filtered and concentrated in vacuo.

Example 3. Obtaining (Ρ) -9- | 4- (Ν-ί.6ζ-Ρvalyloxy) -2- (stearoyloxymethyl) butyl] guanine.

A solution of I-C ^ -B-valine (169 g, 0.67 mol) in dry THF (750 ml) was prepared in a 2 liter flask equipped with a mechanical stirrer, thermometer and dropping funnel. A solution of dicyclohexylcarbodiimide (69.3 g, 0.34 mol) in THF (250 ml) was added over 5 minutes, and the resulting suspension was stirred at 20 ± 5 ° C for 2 hours. Suspension

filtered and the filter cake washed with THF (300 ml). The filtrate and washings were charged into a 3 liter flask equipped with a stirrer and thermometer. The product of example 2 (116 g, 0.22 mol) was added as a solid, rinsing with a THF reactor (250 ml). 4-Щ, I-dimethylamino) pyridine (2.73 g, 0.022 mol) was added and the white slurry was stirred at 20 ± 5 ° C. In 15 minutes, the solids were completely dissolved, and the reaction was completed in 1 h (as judged by HPBC data: column 4.6 x 250 mm 2xBax PxC'8; 85:15 acetonitrile - 0.2% water. HC1O ₄ , addition of at a rate of 1 ml / min; UV detection at 254 nm; the starting material elutes after 4.1 minutes, and the reaction product after 5.9 minutes). The reaction was quenched by the addition of water (5 ml) and the resulting solution was concentrated in vacuo, leaving a light yellow semi-solid. This material was transferred to methanol (1.5 L) and heated to reflux for 30 minutes. The resulting solution was cooled to 25 ° C and the precipitate was removed by filtration. The filtrate was concentrated in vacuo, leaving a viscous, pale yellow oil. Acetonitrile (1 L) was added and the resulting white suspension was stirred at 20 ± 5 ° C for 90 minutes. The crude solid was isolated by filtration, washed with acetonitrile (2 x 100 ml) and dried in air overnight to give the desired product as a waxy, sticky solid (122 g). This material was purified by recrystallization from ethyl acetate (500 ml) and dried in vacuo at 30 ° C to give the desired product as a white, waxy solid (104 g).

Example 4. Obtaining (B) -9- [4- (bivalyloxy) -2- (stearoyloxymethyl) butyl] guanine.

The solution obtained in example 3 (77 g) in warm (40 ° C) ethanol (2.3 L) was loaded into a hydrogenation reactor together with 5% Pb-C (15.4 g). The resulting mixture was stirred at 40 ° C under hydrogen pressure of 40 pounds / ⁱⁿ² (2.8 kg / cm ²⁾ for 4 hours, evacuated and hydrogenated for an additional 4-10 hours. The catalyst was removed by filtration and the filtrate was concentrated under vacuum to removal of white solid. This material was stirred with ethanol (385 ml) at 45 ° C for 1 h, then cooled to 0 ° C and filtered. The filter cake was dried with air, then in vacuum at 35 ° C to obtain the target compound in the form of a white powder (46 g).

Example 5. (K.) - 9- [2- (L-valyloxymethyl) -4 (stearoyloxy) butyl] guanine.

a) (K.) - 9- [2- (I-Boc-b-valiloximethyl) -4 (stearoyloxy) butyl] guanine.

A mixture of Вос-Boc-L-valine (528 mg; 2.1 mmol) and Ν, Ν'-dicyclohexylcarbodiimide (250 mg; 1.21 mg) in dichloromethane (20 ml) was stirred overnight at room temperature, the dicyclohexylurea was filtered and it was extracted with a small volume of dichloromethane, and the filtrate was evaporated in vacuo to a small volume. (B) -9- [2-hydroxymethyl-4 (stearoyloxy) butyl] guanine (340 mg; 0.654 mmol), 4-dimethylaminopyridine (25 mg; 0.205 mmol) and dry Ν, Ν-dimethylformamide (15 ml) were added and the resulting the mixture was stirred for 4 hours at 50 ° C. under nitrogen. The solvent was evaporated in vacuo to a small volume. Column chromatography on silica gel and then on alumina (eluent ethyl acetate: methanol: water in a ratio of 15: 2: 1) gave 185 mg (39%) of the pure target compound as a white solid.

'H NMR spectrum (SPS1 ₃ ) δ: 0.85-1.0 (m, 9H) 18-CH ₃ , CH (CH ₃ ) ₂ ; 1.25 (s, 28H) 4-17-CH ₂ ; 1.44 (s, 9H) t-Vi; 1.60 (quintet, 2H) 3-CH ₂ ; 1.74 (q, 2H) 3'-CH ₂ ; 2.14 (m, 1H) 2'-CH; 2.29 (t, 2H) 2-CH ₂ ; 2.41 (m, 1H) CH (CH ₃ ) ₂ ; 4.1-4.3 (m, 6H) C1'-CH ₂ , C2 '' -CH ₂ , C4-CH ₂ ; 5.4 (d, 1H) aCH; 6.6 (broad s, 2H) guaNH ₂ ; 7.73 (s, 1H) guaN8; 12.4 (wide s).

Spectrum ¹³ C NMR (SPS1 ₃ ) δ: 13.9 (C18); 17.5 / 18.9 (2 Va1 CH ₃ ); 22.4 (C17); 24.7 (C3); 21.8 (C3 '); 28.9-29.3 (C4-6, C15); 29.4 (C7-14); 30.7 (Va1 BC); 31.7 (C16); 34.0 (C2); 35.9 (C2 '); 43.9 (C1 '); 58.7 / Va1 aC); 61.4 / 63.6 (C4 ', C2``); 79.9 (CMe ₃ ); 116.4 (huaC5); 137.9 (huaC7); 151.7 (huaC4); 153.7 (huaC2); 155.7 (СО1МН); 158.8 (huaC6); 172.1 (SSSOO); 173.5 (CH ₂ COO).

B) (B) -9- [2- (b-B aliloxymethyl) -4-stearoloxy) butyl] guanine.

Chilled trifluoroacetic acid (2.0 g) was added to (B) -9- [2-Sch-Boc-Bivaloximethyl) -4- (stearoyloxy) butyl] guanine (180 mg; 0.25 mmol) and the resulting solution was kept at room temperature temperature for 1 h, evaporated to a small volume and lyophilized twice with dioxane to obtain a white amorphous powder. The yield of the target compound obtained in the trifluoroacetate salt was quantitative.

'11 NMR spectrum (ΌΜ8Ο-ά ₆ ) δ: 0.87 (t, 3H) 18-CH ₃ , 0.98 (dd 6H) CH (CH ₃ ) ₂ ; 1.25 (s, 28H) 417-CH ₂ ; 1.50 (quintet, 2H) 3-CH ₂ ; 1.68 (q, 2H) 3'-CH ₂ ; 2.19 (m, 1H) 2'-CH; 2.26 (t, 2H) 2-CH ₂ ; 2.40 (m, 1H) CH (CH ₃ ) ₂ ; 3.9-4.25 (m, 7H) C1'-CH ₂ , C2 '' - CH ₂ , C4-CH ₂ , aCH; 6.5 (broad s, 2H) guaYaH2; 7.79 (s, 1H) guaN8; 8.37 (broad s, 3H) ΝΉ ₃ ⁺ ; 10.73 (broad s, 1H) guaN.

¹³ H NMR Spectrum (ΌΜ8Ο-ά ₆ ) δ: 14.2 (C18); 17.9 / 18.3 (2 Ya1 CH ₃ ); 22.3 (C17); 24.6 (C3); 27.7 (C3 '); 28.7-29.1 (C4-6, C15); 29.2 (C7-14); 29.5 (Va1 βϋ); 31.5 (C16); 33.7 (C2); 35.0 (C2 '); 44.1 (C1 '); 57.6 (Va1 aC); 61.6 / 65.2 (C4 ', C2``); 116.1 (huaC5); 116.3 (hua, 1,290 Hz, CP ₃ ); 137.9 (huaC7);

151.5 (huaC4); 154.0 (huaC2); 156.7 (huaC6); 158.3 (hua, 1 15 Hz, CP ₃ COO) 169.1 (SSSOO); 173.1 (CH2COO).

Example 6. (B) -9- [2-stearoyloxymethyl) -4 (b-valyloxy) butyl] guanine monohydrochloride.

The product of example 3 (360 mg, 0.479 mmol) was dissolved in a mixture of methanol (10 ml) and ethyl acetate (10 ml). 10% Pb / C (100 mg) and ΙΝ HC1 (520 μl) were added to the solution. The reaction mixture was stirred at room temperature for 2 hours at 1 atm. H ₂ . The reaction mixture was filtered and the solvent was evaporated from the filtrate to obtain the desired product as a crystalline solid (300 mg).

Prescription Example A.

Tablet preparation

The following ingredients were sieved through a 0.15 mm sieve and mixed in the dry state:

g (B) -9- [2- (stearoyloxymethyl) -4- (bivalyloxy) butyl] guanine g lactose g crystalline cellulose g magnesium stearate

A tablet machine was used to compress the mixture into tablets containing 250 mg of the active ingredient.

Prescription Example B.

Enteric Coated Tablet

Prescription Example A tablets were spray coated in a coating machine with a solution containing

120 g ethyl cellulose g propylene glycol g sorbitan monooleate in 1000 ml of distilled water. Prescription Example C.

Drug with controlled release of g (B) -9- [2- (stearoyloxymethyl) -4- (bivalyloxy) butyl] guanine, g of hydroxypropylmethyl cellulose (Me11use11 K15),

4.5 g of lactose was mixed in the dry state and granulated with an aqueous povidone paste. Magnesium stearate (0.5 g) was added and the mixture was extruded in a tablet machine into 13 mm tablets containing 500 mg of active agent.

Prescription Example Ό.

Soft capsules

250 g B) -9- [2- (stearoyloxymethyl) -4- (bivalyloxy) butyl] guanine

100 g lecithin

100 g peanut butter

The active ingredient was dispersed in lecithin and peanut butter and soft gelatin capsules were filled with the resulting mixture.

Biological example 1.

Rat Bioavailability Testing

The bioavailability of the compound of formula получ obtained from the intermediate of the present invention was compared with the related compound H2C and derivatives of H2C in a rat model. The compounds of the invention and related compounds were administered orally (using a catheter inserted into the stomach) on groups of three individually weighed animals, each receiving 0.1 mmol / kg of dissolved prodrug in an aqueous solution, based on peanut butter or propylene glycol carrier, depending on the solubility of the active ingredient of the test compound. Animals were deprived of food from 5 hours before use until about 17 hours after use and were kept in metabolic cells. Urine was collected within 24 hours after drug administration and was frozen until analysis. Н2С was analyzed in the urine using HPC / UV analysis according to the method 31aB1e, OEgd, AuBt1stLdei18 CBeto1Bet, 36 No. 2, 339-342 (1992), modified as follows: samples after thawing were diluted in a ratio of 1: 100 with distilled Н ₂ О and filtered through the Amicon filter during centrifugation at a speed of 3000 rpm. within 10 minutes Duplicated 30 μl samples were chromatographed on an HPLC column; 2otax ST-C18; 75-4.6 mm; 3.5 microns; the mobile phase is 0.05 M ΝΗ ₄ , ₄ , 3-4% methanol, pH 3.3-3.5; 0.5 ml / min .; 254 nm, the retention time of H2C at a concentration of MeOH4% and a pH of 3.33 - 12.5 minutes Bioavailability was calculated as the measured H2C excretion from each animal, averaged from at least three animals and expressed as the percentage of the averaged 24-hour excretion of H2C from urine from a group of 4 individually weighed rats, respectively, receiving intravenous injections of 0.1 mmol / kg H2C in Ringer's buffer solution, performing the analysis as described above.

The compound in comparative examples was obtained analogously to those described in this application.

The results are shown in the following table. 2.

table 2

Compound K2 K2 Bioavailability Comparative Example 1 Hydrogen Hydrogen 8% Reference Example 2 Felled Hydrogen 29% Reference Example 3 Felled Felled 36% Example of the invention Felled Stearoyl 56% Reference Example 4 Stearoyl Stearoyl one% Example of the invention Felled Myristoyl 57% Example of the invention Felled Oleil 51% Example of the invention Felled Butyryl 45% Reference Example 5 Felled Acetyl eleven% Example of the invention Felled Decanoyl 48% Example of the invention Felled Docosanoil 48%

Example of the invention Isoleucil Stearoyl 53% Example of the invention Isoleucil Decanoyl 57% Example of the invention Isoleucil Myristoyl 49% Example of the invention Felled 4-acetyl butyryl 52% Example of the invention Felled Dodecanoyl 46% Example of the invention Felled Palmitoyl 58% Example of the invention Stearoyl Felled 52% Reference Example 6 Alanine Stearoyl 23% Reference Example 7 Glycyl Decanoyl 25% Reference Example 8 Acetyl Felled 7% Reference Example 9 Hydrogen Stearoyl 12% Reference Example 10 Stearoyl Hydrogen 7%

Comparison of the bioavailability of the compounds of formula I with comparative examples shows that a specific combination of fatty acids according to K. | / K. ₂ with amino acids of B ₁ / B ₂ provides significantly higher bioavailability values than the corresponding diamino acid ester or dibasic fatty acid ester. For example, within the framework of such a model, the compound according to the invention - (K.) - 9- [2- (steryloxymethyl) -4- (b-valyloxy) butyl] guanine demonstrates 55% better bioavailability than the corresponding diviline ester Comparative Example 3, or, respectively, - (B) 9- [2- (butyryloxymethyl) -4- (b-valyloxy) butyl] guanine demonstrates 25% better bioavailability than the corresponding diviline ester.

It is also quite obvious, for example, from comparative examples 5, 6 and 7, that only the special fatty acids of the present invention in combination with specific amino acids provide such an unexpected increase in pharmacokinetic parameters.

Biological example 2.

Plasma concentrations in rats

Plasma concentration analysis was performed on male rattadia Ea \\ '1eu rats. Animals were deprived of food during the night before the introduction of the drug, but provided free access to water. Each of the test compounds was prepared as a solution / suspension in propylene glycol with a concentration corresponding to 10 mg H2C / ml and shaken at room temperature for eight hours. Groups of rats (at least 4 rats in each group) received an oral dose of 10 mg / kg (1 ml / kg) of each of the compounds; each dosage was administered through a gastric tube. At certain intervals after taking the dose (0.25, 0.5, 1, 1.5, 2, 4, 6, 9, 12, 15 and 24 hours after dosing), heparinized blood samples were obtained from the tail vein of each animal ( 0.4 ml / sample). These blood samples were immediately cooled in an ice bath. Within two hours of collection, the plasma was separated from the red blood cells by centrifugation and frozen until further analysis. The components of interest were separated from plasma proteins by precipitation in acetonitrile. After lyophilization and reconstruction, plasma concentrations were determined by reversed-phase HPLC with fluorescence detection. The oral absorption of H2C and other test compounds was determined by comparing the area of H26 under the curve for the oral dosage with the corresponding value obtained by administering an intravenous dose of H2C of 10 mg / kg, which was used on a separate group of rats. The results are presented in the above table. 1B.

Biological Example 3. Biological Applicability in Monkeys The compounds of the invention and comparative example 3 (see biological example 1 above) used p. about. through a stomach tube on monkeys supoto1sch§. The solutions used contained

The active ingredient according to 150 mg, dissolved in 6.0 invention, is (K) -9- [2- (steroylpropylene glycol, which is coxymethyl) -4- (b-valyloxy) is 25 mg / kg or tyl] guanine 0, 0295 mmol / kg

Comparative example 3 164 mg was dissolved in 7.0 ml of water, which corresponds to 23.4 mg / kg or 0.0295 mmol / kg

Blood samples were taken after 30 minutes, 1, 2, 3, 4, 6, 10 and 24 hours. Plasma was separated by centrifugation at a speed of 2500 rpm. and the samples were inactivated at 54 ° C for 20 minutes before being frozen until subsequent analyzes. Plasma H2C levels were recorded using HPLC / UV analysis as described above in Example 30.

In FIG. 1 shows the release of H26 plasma as a function of time. Although it is not possible to draw statistically significant conclusions from single animal experiments, it can be assumed that animals treated with the compound of the present invention experience, to some extent, a more rapid and stronger effect of H2C. than animals that used the alternative H26 prodrug.

Biological example 4.

Antivirus activity

Mice infected with herpes simplex virus-1 (H8U-1) served as a model for determining the efficacy of antiviral agents ίη U1uo. On mice inoculated intraperitoneally with H8U-1, at a 1000-fold LIBO value, a formulation containing the currently available anti-herpes agent acyclovir (21 and 83 mg / kg in 2% propylene glycol in a vehicle containing sterile water three times a day, orally) was used ), or the compound of Example 29 (21 and 83 mg / kg in 2% propylene glycol, in a vehicle containing sterile water, three times a day, orally), for 5 consecutive days, starting to be used 5 days after inoculation. Animal mortality was estimated daily. The results are shown in FIG. 2, which shows the dependence of the degree of survival on time. In the caption to the figure, the compound of the invention is designated as Ex.29, and acyclovir is designated as ACS. The percentage of mice surviving H8U-1 infection turned out to be significantly greater when using this dosage of a compound of the invention, compared to using an equivalent dose of acyclovir.

The above material is for illustration only, and the present invention is not limited to the content of this material. It is understood that various variations and changes that are obvious to a person skilled in this field are covered by the scope of the claims of the invention, as reflected in the attached claims.

Claims (4)

CLAIM 1. The compound of the formula in which K ₆ and K ₇ are _C1-7 alkyl or benzyl, or K ₆ and K ₇ together are —CH ₂ CH ₂ - or —CH ₂ CH ₂ CH ₂ -, —CH ₂ CH ₂ CH ₂ CH ₂ - and K ₉ represents hydrogen or an alcohol protecting group. 2. The compound of claim 1, wherein K ₉ is hydrogen. 3. The compound according to claim 1, in which the alcohol protecting group is a benzyl group. 4. The compound according to claim 1 or 2, in which K ₆ and K ₇ each is ethyl.